Pyrosequencing
Overview
DNA sequencing
DNA sequencing includes several methods and technologies that are used for determining the order of the nucleotide bases—adenine, guanine, cytosine, and thymine—in a molecule of DNA....
(determining the order of nucleotides in DNA) based on the "sequencing by synthesis
Polymerase chain reaction
The polymerase chain reaction is a scientific technique in molecular biology to amplify a single or a few copies of a piece of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence....
" principle. It differs from Sanger sequencing, in that it relies on the detection of pyrophosphate
Pyrophosphate
In chemistry, the anion, the salts, and the esters of pyrophosphoric acid are called pyrophosphates. Any salt or ester containing two phosphate groups is called a diphosphate. As a food additive, diphosphates are known as E450.- Chemistry :...
release on nucleotide incorporation, rather than chain termination with dideoxynucleotides
Dideoxynucleotides
Dideoxynucleotides, or ddNTPs, are nucleotides lacking a 3'-hydroxyl group on their deoxyribose sugar. Since deoxyribose already lacks a 2'-OH, dideoxyribose lacks hydroxyl groups at both its 2' and 3' carbons...
. The technique was developed by Pål Nyrén
Pål Nyrén
Pål Nyrén is a biochemistry professor at the Royal Institute of Technology , Stockholm. He is most famous for developing the pyrosequencing method for DNA sequencing.-Career:*1999 Professor in Biochemistry, KTH, Stockholm...
and Mostafa Ronaghi
Mostafa Ronaghi
Mostafa Ronaghi, born 1968. is a molecular biologist, specializing in DNA sequencing methodology . He earned his Ph.D. from the Royal Institute of Technology in Sweden in 1998.He is currently the Chief Technology Officer and Senior Vice President at Illumina...
at the Royal Institute of Technology
Royal Institute of Technology
The Royal Institute of Technology is a university in Stockholm, Sweden. KTH was founded in 1827 as Sweden's first polytechnic and is one of Scandinavia's largest institutions of higher education in technology. KTH accounts for one-third of Sweden’s technical research and engineering education...
in Stockholm in 1996.
"Sequencing by synthesis" involves taking a single strand of the DNA to be sequenced and then synthesizing its complementary strand enzymatically.
Discussions